摘要
本室以前已经报道G蛋白偶联受体APJ的内源性配体多肽apelin-13促进单核细胞-血管内皮细胞黏附,本文研究PI3K信号途径是否参与apelin-13促进单核细胞-血管内皮细胞黏附,探讨apelin/APJ系统的细胞信号转导机制.MPO方法检测细胞黏附;Western blot方法检测PI3K、VCAM-1的表达.Western blot方法结果显示,apelin-13(0、0.5、1、2、4μmol/L)浓度依赖性刺激血管内皮细胞PI3K磷酸化,以1μmol/L最为明显;1μmol/L apelin-13时间依赖性促进血管内皮细胞PI3K磷酸化,在30 min增加最为显著;PI3K抑制剂LY294002明显抑制apelin-13诱导的VCAM-1表达和单核细胞-血管内皮细胞黏附.上述结果表明,PI3K信号途径介导apelin-13促进单核细胞-血管内皮细胞黏附.
Previously we reported that G protein-coupled receptor APJ endogenous ligand apelin-13 induced adhesion of monocytes to human umbilical vein endothelial cells（HUVECs）.Now we investigated whether phosphatidylinositol 3-kinase（PI3K） signaling pathway mediated monocytes（MCs） adhesion to HUVECs induced by apelin-13.Human umbilical vein endothelial cell line ECV304 was cultured in DMEM medium and the monocyte cell line THP-1 was cultured in 1640 medium.Myeloperoxidase（MPO） assay was used to identify effects of monocytes adhesion to HUVECs.Expression of vascular cell adhesion molecule（VCAM）-1,phospho-PI3K and PI3K were detected by Western blotting.Apelin-13 promoted PI3K phosphorylation in concentration-dependent and time-dependent manner,which reached the peak at 1 μmol/L and 30 min respectively.The PI3K inhibitor LY294002 inhibited PI3K phosphorylation and the expression of VCAM-1 in HUVECs induced by apelin-13.And the PI3K inhibitor LY294002 inhibited the MCs adhesion to HUVECs induced by apelin-13.It can be concluded that apelin-13 promoted monocytes adhesion to HUVECs through VCAM-1 mediated by PI3K signaling pathway.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2011年第12期1162-1170,共9页
Progress In Biochemistry and Biophysics
基金
supported by grants from The National Natural Science Foundation of China(30901577,30770868)
National Basic Research Program of China(2006CB503808)
Talent Program of USC(2008XQD01)
Hengyang Municipal Bureau of Science and Technology(2009KJ14)
Ministry of Education of Returned Overseas Students to Start Research and Fund Projects(20091590)
Aid Program for Science and Technology Innovative Research Team in Higher Educational Institutions of Hunan Province~~
